(R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents

N. R. DeStefino, A. A. Pilato, M. Dittrich, S. V. Cherry, S. Cho, J. R. Stiles, S. D. Meriney

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

(R)-roscovitine (Ros) is a cyclin-dependent kinase inhibitor that also has been shown to have direct agonist and antagonist actions on Cav2.1 (P/Q-type) and Cav 2.2 (N-type) families of voltage-gated calcium channels. These kinase-independent effects represent a novel opportunity to advance our understanding of calcium channel function and calcium-triggered neurotransmitter release. Furthermore, such actions on calcium channels may direct the development of Ros derivatives as new therapeutic agents. We used patch clamp recordings to characterize mechanisms that underlie the agonist effects of Ros on unitary N-type calcium channel gating. We found that N-type channels normally gate with either a short or long mean open time, that Ros significantly prolonged the mean open time of the long gating component and increased the probability of observing channels that gated with a long open time, but had no effect on single channel conductance. Using Monte Carlo simulations of a single channel kinetic model and Ros interactions, we were able to reproduce our experimental results and investigate the model's microscopic dynamics. In particular, our simulations predicted that the longer open times generated by Ros were due to the appearance of a long open state combined with an increased amount of time spent in transitions between open states. Our results suggest a mechanism for agonist effects of Ros at the level of single channels, and provide a mechanistic explanation for previously reported agonist effects on whole cell calcium currents.

Original languageEnglish (US)
Pages (from-to)838-849
Number of pages12
JournalNeuroscience
Volume167
Issue number3
DOIs
StatePublished - May 1 2010

Fingerprint

N-Type Calcium Channels
Calcium Channels
Calcium
Cyclin-Dependent Kinases
roscovitine
Neurotransmitter Agents
Phosphotransferases

Keywords

  • Channel kinetics
  • Conductance
  • Patch clamp
  • Roscovitine
  • Single channel current
  • Voltage-gated calcium channel

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

DeStefino, N. R., Pilato, A. A., Dittrich, M., Cherry, S. V., Cho, S., Stiles, J. R., & Meriney, S. D. (2010). (R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents. Neuroscience, 167(3), 838-849. https://doi.org/10.1016/j.neuroscience.2010.02.041

(R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents. / DeStefino, N. R.; Pilato, A. A.; Dittrich, M.; Cherry, S. V.; Cho, S.; Stiles, J. R.; Meriney, S. D.

In: Neuroscience, Vol. 167, No. 3, 01.05.2010, p. 838-849.

Research output: Contribution to journalArticle

DeStefino, NR, Pilato, AA, Dittrich, M, Cherry, SV, Cho, S, Stiles, JR & Meriney, SD 2010, '(R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents', Neuroscience, vol. 167, no. 3, pp. 838-849. https://doi.org/10.1016/j.neuroscience.2010.02.041
DeStefino, N. R. ; Pilato, A. A. ; Dittrich, M. ; Cherry, S. V. ; Cho, S. ; Stiles, J. R. ; Meriney, S. D. / (R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents. In: Neuroscience. 2010 ; Vol. 167, No. 3. pp. 838-849.
@article{2196f628b3d047aa967e1451f1ba6556,
title = "(R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents",
abstract = "(R)-roscovitine (Ros) is a cyclin-dependent kinase inhibitor that also has been shown to have direct agonist and antagonist actions on Cav2.1 (P/Q-type) and Cav 2.2 (N-type) families of voltage-gated calcium channels. These kinase-independent effects represent a novel opportunity to advance our understanding of calcium channel function and calcium-triggered neurotransmitter release. Furthermore, such actions on calcium channels may direct the development of Ros derivatives as new therapeutic agents. We used patch clamp recordings to characterize mechanisms that underlie the agonist effects of Ros on unitary N-type calcium channel gating. We found that N-type channels normally gate with either a short or long mean open time, that Ros significantly prolonged the mean open time of the long gating component and increased the probability of observing channels that gated with a long open time, but had no effect on single channel conductance. Using Monte Carlo simulations of a single channel kinetic model and Ros interactions, we were able to reproduce our experimental results and investigate the model's microscopic dynamics. In particular, our simulations predicted that the longer open times generated by Ros were due to the appearance of a long open state combined with an increased amount of time spent in transitions between open states. Our results suggest a mechanism for agonist effects of Ros at the level of single channels, and provide a mechanistic explanation for previously reported agonist effects on whole cell calcium currents.",
keywords = "Channel kinetics, Conductance, Patch clamp, Roscovitine, Single channel current, Voltage-gated calcium channel",
author = "DeStefino, {N. R.} and Pilato, {A. A.} and M. Dittrich and Cherry, {S. V.} and S. Cho and Stiles, {J. R.} and Meriney, {S. D.}",
year = "2010",
month = "5",
day = "1",
doi = "10.1016/j.neuroscience.2010.02.041",
language = "English (US)",
volume = "167",
pages = "838--849",
journal = "Neuroscience",
issn = "0306-4522",
publisher = "Elsevier Limited",
number = "3",

}

TY - JOUR

T1 - (R)-roscovitine prolongs the mean open time of unitary N-type calcium channel currents

AU - DeStefino, N. R.

AU - Pilato, A. A.

AU - Dittrich, M.

AU - Cherry, S. V.

AU - Cho, S.

AU - Stiles, J. R.

AU - Meriney, S. D.

PY - 2010/5/1

Y1 - 2010/5/1

N2 - (R)-roscovitine (Ros) is a cyclin-dependent kinase inhibitor that also has been shown to have direct agonist and antagonist actions on Cav2.1 (P/Q-type) and Cav 2.2 (N-type) families of voltage-gated calcium channels. These kinase-independent effects represent a novel opportunity to advance our understanding of calcium channel function and calcium-triggered neurotransmitter release. Furthermore, such actions on calcium channels may direct the development of Ros derivatives as new therapeutic agents. We used patch clamp recordings to characterize mechanisms that underlie the agonist effects of Ros on unitary N-type calcium channel gating. We found that N-type channels normally gate with either a short or long mean open time, that Ros significantly prolonged the mean open time of the long gating component and increased the probability of observing channels that gated with a long open time, but had no effect on single channel conductance. Using Monte Carlo simulations of a single channel kinetic model and Ros interactions, we were able to reproduce our experimental results and investigate the model's microscopic dynamics. In particular, our simulations predicted that the longer open times generated by Ros were due to the appearance of a long open state combined with an increased amount of time spent in transitions between open states. Our results suggest a mechanism for agonist effects of Ros at the level of single channels, and provide a mechanistic explanation for previously reported agonist effects on whole cell calcium currents.

AB - (R)-roscovitine (Ros) is a cyclin-dependent kinase inhibitor that also has been shown to have direct agonist and antagonist actions on Cav2.1 (P/Q-type) and Cav 2.2 (N-type) families of voltage-gated calcium channels. These kinase-independent effects represent a novel opportunity to advance our understanding of calcium channel function and calcium-triggered neurotransmitter release. Furthermore, such actions on calcium channels may direct the development of Ros derivatives as new therapeutic agents. We used patch clamp recordings to characterize mechanisms that underlie the agonist effects of Ros on unitary N-type calcium channel gating. We found that N-type channels normally gate with either a short or long mean open time, that Ros significantly prolonged the mean open time of the long gating component and increased the probability of observing channels that gated with a long open time, but had no effect on single channel conductance. Using Monte Carlo simulations of a single channel kinetic model and Ros interactions, we were able to reproduce our experimental results and investigate the model's microscopic dynamics. In particular, our simulations predicted that the longer open times generated by Ros were due to the appearance of a long open state combined with an increased amount of time spent in transitions between open states. Our results suggest a mechanism for agonist effects of Ros at the level of single channels, and provide a mechanistic explanation for previously reported agonist effects on whole cell calcium currents.

KW - Channel kinetics

KW - Conductance

KW - Patch clamp

KW - Roscovitine

KW - Single channel current

KW - Voltage-gated calcium channel

UR - http://www.scopus.com/inward/record.url?scp=77951296911&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951296911&partnerID=8YFLogxK

U2 - 10.1016/j.neuroscience.2010.02.041

DO - 10.1016/j.neuroscience.2010.02.041

M3 - Article

C2 - 20188151

AN - SCOPUS:77951296911

VL - 167

SP - 838

EP - 849

JO - Neuroscience

JF - Neuroscience

SN - 0306-4522

IS - 3

ER -